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  • Combination of TransPass D1 or TransPass D2 & TransPass V (M2561)

    Protocol

    1. In a tube, mix plasmid(s) and/or siRNA in serum free medium.
    2. Add TransPass D1 or TransPass D2 to the above mixture.
    3. Incubate at room temperature for 25-30 minutes.
    4. Add TransPass V to the incubated mixture.
    5. (Optional) Incubate at room temperature for 15 minutes.
    6. Add the transfection complex mixture to cells (in complete growth media).
    7. Incubate at 37°C, 5% CO2 for 24 hours before replacing media. 
      Note: Replace media every day, if longer incubation period is required. 

      Table 1:
      Transfection complex mixtures using TransPass D1 or TransPass D2 & TransPass V
      Culture Vessel Surface
      Area
      Volume
      of Plating
      Medium
      (per well)
      *Total DNA in
      Serum-free
      Medium Volume
      (per well)
      TransPass
      D1 or D2
      (per well)
      TransPass V
      (per well)
      96 well 0.32 cm2 100 µ 0.1 µg in 10 µl 0.1–0.3 µl 0.1–0.9 µl
      48 well 0.95 cm2 250 µ 0.3 µg in 25 µl 0.3–0.9 µl 0.3–2.7 µl
      24 well 1.9 cm2 500 µl 0.7 µg in 50 µl 0.7–2.1 µl 0.7–6.3 µl
      12 well 3.8 cm2 1 ml 1.5 µg in 150 µl 1.5-4.5 µl 1.5–13.5 µl
      35 mM or 6 well 9.5 cm2 2 ml 3 µg in 250 µl 3–9 µl 3–27 µl
      60 mM dish 21 cm2 5 ml 6 µg in 500 ml 6–18 µl 6–54 µl
      100 mM dish 55 cm2 15 ml 18 µg in 1 ml 18–54 µl 18–162 µl<
      *For plasmid and siRNA transfection, we recommend 20–100 nM siRNA per well in addition to the suggested DNA amount and the 1:1, 1:2 or 1:3 ratio (TransPass transfection reagent: TransPass V).